CN102234329A - Plant-type associated and/or yield associated protein, coding gene thereof, and application thereof - Google Patents

Abstract

The invention discloses a plant-type associated and/or yield associated protein, a coding gene thereof, and an application thereof. The protein provided by the invention is a protein showed by the following description (a) or (b): (a) a protein composed of an amino acid sequence represented by SEQ ID NO: 2; (b) a protein formed after an amino acid sequence represented by SEQ ID NO: 2 is processed after substitutions and/or deletions and/or additions of one or more amino acid residues, wherein the obtained protein is derived from the protein represented by (a), and is plant-type associated and/or yield associated. As a result of experiments, when the gene provided by the invention is introduced into a paddy mutant nall, a plant type of a transgenic plant can be substantially improved. Specifically, the improvements are represented in the following respects that: a main stem height is larger than that of the mutant nall, a main panicle length is larger than that of the mutant mall, a main panicle grain amount is larger than that of the mutant mall, a leaf width is larger than that of the mutant mall, and a leaf length is larger than that of the mutant mall,. Paddy rice yield of a plant with improved plant type can be increased. The gene provided by the invention has a good application prospect in fields of plant genetic breeding and crop yield improving.

Description

A kind of relevant with plant type and/or with output associated protein and encoding gene and application

Technical field

The present invention relates to a kind of relevant with plant type and/or with output associated protein and encoding gene and application.

Background technology

Plant type (plant type) is meant stack features relevant with crop varieties output ability or plant materials at the spatial arrangement mode, i.e. the growing way appearance.Ideotype (Ideal plant type) also is called ideal type (Ideotype), finger by favourable plant photosynthesis, grow and idealized plant type that the proterties of grain yield is formed, it can improve colony's efficiency of light energy utilization to greatest extent, increases biological yield and improves economic coefficient etc.The research of relevant plant type of rice, phase late 1950s, Japan scientist angle Tian Chongsan youth sums up the kind that is suitable for how fertile intensive culture and should have thick, little, upright and bottle-green blade from his research practice to paddy rice, soybean, sugarcane etc., short and the tough and tensile stem stalk and the ideotype theory of medium ability for tillering (angle Tian Chongsan youth. agricultural and gardening .1987,62 (1): 25～29.).Constantly the improvement plant type also is the basic experience of China's super high-yielding rice breeding.Drape over one's shoulders blade profile from the high stalk of farm variety and progressively improve and be straight blade profile of short stem, the form improvement develops into form gradually and the function improvement is laid equal stress on from paying attention to, and has not only improved optical energy utilization efficiency, has increased lodging tolerance, has also strengthened the physiological function of paddy rice.

The rice leaf proterties is the important factor that plant type constitutes, be directly connected to the photosynthetic area and the efficiency of light energy utilization of blade, and then to output generation material impact, therefore the research to blade profile is one of breeding man, geneticist and molecular biologist Focal Point of Common Attention, and the width of blade and curling situation are important component parts wherein.Studies show that paddy rice narrow leaf and leaf roll proterties are controlled by qualitative trait gene mainly.

Summary of the invention

An object of the present invention is to provide a kind of albumen and encoding gene thereof.

Albumen provided by the present invention, be following a) or b) protein:

A) protein of forming by the aminoacid sequence shown in the SEQ ID NO:2;

B) with the aminoacid sequence shown in the SEQ ID NO:2 through the replacement of one or several amino-acid residue and/or disappearance and/or interpolation and relevant with the plant plant type and/or be correlated with output by a) deutero-protein.

Described proteic encoding gene is following 1), 2), 3) or 4) gene:

1) its nucleotide sequence is from dna molecular shown in the Nucleotide of 5 ' terminal 140-1201 position among the SEQ ID NO:1;

2) its nucleotide sequence is a dna molecular shown in the SEQ ID NO:1;

3) under stringent condition with 1) or 2) the dna sequence dna hybridization that limits and the dna molecular of encoding said proteins;

4) with 1) or 2) dna sequence dna that limits has the homology more than 90% and the dna molecular of encoding said proteins.

The rigorous condition of described height be 0.1 * SSPE (or 0.1 * SSC), in the solution of 0.1%SDS, hybridization and wash film under 65 ℃ of conditions.

In order to make the albumen in (a) be convenient to purifying, proteinic N-terminal or C-terminal that can the aminoacid sequence shown in the SEQ ID NO:2 is formed in by sequence table connect label as shown in table 1.

The sequence of table 1 label

Label	Residue	Sequence
			Poly-Arg	5-6 (being generally 5)	RRRRR
Poly-His	2-10 (being generally 6)	HHHHHH
			FLAG	8	DYKDDDDK
Strep-tag?II	8	WSHPQFEK
			c-myc	10	EQKLISEEDL

Above-mentioned (a) but in the albumen synthetic, also can synthesize its encoding gene earlier, carry out biology again and express and to obtain.Proteic encoding gene in above-mentioned (a) can be by the codon with one or several amino-acid residue of disappearance in the dna sequence dna shown in the SEQ ID NO:1, and/or carry out the missense mutation of one or several base pair, and/or obtain at the encoding sequence that its 5 ' end and/or 3 ' end connects the label shown in the table 1.

Increase above-mentioned arbitrary described encoding gene total length or its any segmental primer to also belonging to protection scope of the present invention.

A primer sequence of described primer centering is shown in SEQ ID NO:3, and another primer sequence of described primer centering is shown in SEQ ID NO:4.

The recombinant vectors, reorganization bacterium, transgenic cell line or the expression cassette that contain above-mentioned arbitrary described encoding gene also belong to protection scope of the present invention.

Described recombinant vectors is that the multiple clone site that above-mentioned arbitrary described encoding gene inserts carrier pCAMBIA2300-Actin is obtained.

For the ease of transgenic plant cells or plant being identified and screening, can process used plant expression vector, can produce the enzyme of colour-change or the gene of luminophor (gus gene, luciferase genes etc.) as adding the coding that in plant, to express, have the antibiotic marker thing (gentamicin marker, kantlex marker etc.) of resistance or anti-chemical reagent marker gene (as anti-weedkiller gene) etc.From the security consideration of transgenic plant, can not add any selected marker, directly with adverse circumstance screening transformed plant.

Carry gene of the present invention plant expression vector can Ti-plasmids, Ri plasmid, plant viral vector, directly DNA conversion, microinjection, electricity be led, conventional biological method transformed plant cells or tissue such as agriculture bacillus mediated by using, and the plant transformed cell or tissue is cultivated into plant.

The application of above-mentioned arbitrary described encoding gene in improvement plant plant type and/or improvement output phenotype and/or raising output also belongs to protection scope of the present invention.

The application of above-mentioned arbitrary described albumen in improvement plant plant type and/or improvement output phenotype and/or raising output also belongs to protection scope of the present invention.

Last purpose of the present invention provides a kind of method that improves plant plant type and/or improvement output phenotype and/or improve output.

The method of improvement plant plant type provided by the present invention and/or improvement output phenotype and/or raising output is following 1) or 2) or 3) described:

1) in the plant that sets out, imports claim 2 or 3 described encoding genes, obtain comparing the purpose plant of plant type improvement with the plant that sets out;

2) in the plant that sets out, import claim 2 or 3 described encoding genes, obtain comparing the purpose plant of output phenotype improvement with the plant that sets out;

3) in the plant that sets out, import claim 2 or 3 described encoding genes, obtain comparing the purpose plant of output raising with the plant that sets out.

Above-mentioned arbitrary described encoding gene imports by above-mentioned arbitrary described recombinant vectors.

In above-mentioned arbitrary described application and the above-mentioned arbitrary improvement plant plant type method, described plant type improvement is wider than the plant that sets out for the tall and big plant leaf in the plant that sets out, purpose plant of plant stem of purpose plant, and/or the plant leaf of purpose plant is grown up in the plant that sets out;

The improvement of described output phenotype for the plant master spike length of purpose plant greater than the plant master grain number per spike of set out plant and/or purpose plant more than the plant that sets out.

In above-mentioned arbitrary described application and the above-mentioned arbitrary improvement plant plant type method, the described plant that sets out is a monocotyledons; Described monocotyledons is a paddy rice; Described paddy rice is rice mutant nal1.

Experimental results show that, after in rice mutant nal1, importing gene of the present invention, the plant type of the transfer-gen plant that obtains and output phenotype are significantly improved, and are in particular in: the plant stem is tall and big to be wider than mutant nal1, plant leaf in mutant nal1, plant leaf and to grow up in mutant nal1, plant master spike length greater than mutant nal1, plant master grain number per spike more than mutant nal1.The rice yield of plant after the plant type improvement can improve.Gene of the present invention will have broad application prospects in genetic breeding field, the raising crop yield field of plant.

Description of drawings

Fig. 1 is the genetic map of SUN1mu gene map based cloning.

Fig. 2 is the phenotype of transgenic paddy rice and adjoining tree.

Fig. 3 is that the leaf of transgenic paddy rice and adjoining tree is wide and leaf is long.

Fig. 4 is the phenotype of fringe and the internode of transgenic paddy rice and adjoining tree.

Fig. 5 is the transparent and crosscut comparison diagram in the middle part of transgenic paddy rice and adjoining tree.

Embodiment

Employed experimental technique is ordinary method if no special instructions among the following embodiment.

Used material, reagent etc. if no special instructions, all can obtain from commercial channels among the following embodiment.

The discovery of embodiment 1, gene

Under nal1 (narrow leaf 1) mutant background, screen it with the method for EMS mutagenesis and recover mutant, through 3 generation genetic stability, obtain an obvious regressive mutant of phenotype, called after sun1 (suppressor of nal1).

Adopt the map based cloning method to separate from sun1 and plant type genes involved SUN1mu, concrete grammar is as follows:

1) be material with the F2 that nal1 (japonica rice background) hybridization is obtained for segregating population with sun1 mutant (long-grained nonglutinous rice background), therefrom identified 1600 individualities and carried out genetic analysis, the SUN1mu assignment of genes gene mapping has been cloned on (AC133340) to No. 3 chromosomal BAC of paddy rice with sun1 mutation type surface.

2) utilize long-grained nonglutinous rice (9311), difference in this regional DNA sequence between the japonica rice (Japan is fine) designs the CAPS mark, further target gene is navigated in the scope of a 29KB and (see Fig. 1, the number of recombinant chou between digitized representation molecule marker and goal gene, arrow is illustrated in the insertion that cds begins 1 a between the 214th and 215 base), this dna fragmentation is carried out sequential analysis, discovery has only an encoding gene ORF1, its cDNA has the nucleotide sequence of SEQ ID NO:1, by 1353 based compositions, its encoder block is that coding has the protein of the amino acid residue sequence of SEQ ID NO:2 in the sequence table from 5 ' end 140-the 1201st bit base.

3) utilize the method for order-checking to find transgenation.If this gene is the candidate gene of SUN1mu gene, just should be between sun1 mutant (long-grained nonglutinous rice background) and nal1 (long-grained nonglutinous rice background) representation sequence difference.Concrete grammar is: above-mentioned ORF is carried out also order-checking of pcr amplification, and (detecting primer sequence is: 5 '-GATGGACTCCCCGTCGCCTAT and 5 '-GTGGTGGGAGCTAGAGGCTCA), the result is representation sequence difference between sun1 mutant (long-grained nonglutinous rice background) and nal1 (long-grained nonglutinous rice background); Simultaneously its genomic The sequencing results is shown that ORF1 produces the insertion of 1 base in the sun1 mutant, this sudden change causes from the 73rd amino acids residue phase shift mutation taking place later in the coded amino acid residue sequence of SUN1mu.Therefore, infer that this ORF1 is the candidate gene of SUN1mu.

Thus, the cDNA nucleotide sequence of the gene SUN1mu that obtains is shown in SEQ ID NO:1, and by 1353 based compositions, from its 5 ' end 140-1201 bit base proteins encoded, the aminoacid sequence of encoded protein is SEQ IDNO:2.

The preparation of embodiment 2, gene and functional verification

Wild-type paddy rice Zhejiang spoke 802 (zf802) are at document (Zheng Leiying, Zhu Xudong, before the money, Zhao Zhong, Zhang Jianjun, Hu Xiaohe, Lin Hongxuan, form and the positioning analysis of the Luo Da .2003. paddy rice fringe mutant C1 of portion. Science Bulletin the 48th the 3rd phase of volume) in disclosed, provide by Inst. of Genetics and Development Biology, CAS.

The pMD19-T carrier is available from takara, and catalog number is D102A.

Carrier pCAMBIA2300-Actin is at document (Kejian Wang, Ding Tang, Lilan Hong, WenyingXu, Jian Huang, Ming Li, Minghong Gu, Yongbiao Xue, Zhukuan Cheng.2010.DEPandAFORegulate Reproductive Habit in Rice.January 2010.Volume 6.) disclosed in, provided by Inst. of Genetics and Development Biology, CAS.

Agrobacterium tumefaciens EH105 document (Zhao Zhiqiang, Fu Yaping, poplar Kun, Zhang Yuman, the face Yongsheng, Fang Rongxiang, Sun Zongxiu, the clone and the functional analysis .Chin J Biotech2008 of Chen Xiao English .2008. paddy rice small G-protein OsPra2 gene, December 25; 24 (12): disclosed among the 2027-2033, and provided by Inst. of Genetics and Development Biology, CAS.

Rice mutant nal1 (Dong Fenggao, Xiong Zhenmin, Qian Qian, Zu Xudong, Chen Shihua (1994) .Breeding Near-isogenic lines of morphological markers in indica rice.Chinese J.Rice sci.8,135-139.(providing) by Inst. of Genetics and Development Biology, CAS.

One, gene preparation

From the DNA shown in 5 ' the end 140-1201 bit base, also can be prepared as follows among the SEQ ID NO:1 among the synthetic SEQ ID NO:1 from the DNA shown in 5 ' the end 140-1201 bit base.Hold the proteic aminoacid sequence of the dna encoding shown in the 140-1201 bit base shown in SEQ ID NO:2 from 5 ' among the SEQ ID NO:1.

Extracting total RNA of sun1 mutant, is template with its reverse transcription product, carries out pcr amplification with following primer P1/P2, obtains pcr amplification product.

P1：5’-ATGGACTCCCCGTCGCCTAT(SEQ?ID?NO：3)；

P2：5’-TCACACACGAGGAGGAATTGGA(SEQ?ID?NO：4)。

The PCR product is carried out 1% agarose gel electrophoresis detect, reclaim the also dna fragmentation of purifying 1062bp, be connected, will connect the product transformed into escherichia coli, resistance screening, picking mono-clonal with the pMD-19T carrier; Mono-clonal is carried out liquid culture respectively, extract plasmid, plasmid is checked order.As a result, the gene order of inserting in the pMD-19T carrier as among the SEQ ID NO:1 from shown in 5 ' the end 140-1201 bit base, show that the recombinant vectors of structure is correct, with its called after pMD-19T-SUN1mu.

Two, gene transformation mutant nal1

1, recombinant expression vector makes up

Cut pMD-19T-SUN1mu with restriction enzyme Xba I and Sal I enzyme, reclaim target gene fragment; Cut carrier pCAMBIA2300-Actin with restriction enzyme Xba I and Sal I enzyme, reclaim the big fragment of carrier; Target gene fragment is connected with the big fragment of carrier, obtains recombinant vectors.With the recombinant vectors transformed into escherichia coli, resistance screening, picking mono-clonal; Mono-clonal is carried out liquid culture respectively, extract plasmid, plasmid is checked order.As a result, the gene order of inserting between the Xba I of pCAMBIA2300-Actin carrier and SalI restriction enzyme site as among the SEQ ID NO:1 from shown in 5 ' the end 140-1201 bit base, show that the recombinant vectors of structure is correct, with its called after pActin::SUN1-mu.

2, the reorganization Agrobacterium makes up

With the thermal shock method recombinant expression vector pActin::SUN1-mu is transformed agrobacterium tumefaciens EH105, screen positive recombinant, obtain containing the agrobacterium tumefaciens EH105 of recombinant expression vector pActin::SUN1-mu, note is made EH105-pActin::SUN1-mu.

3, transfer-gen plant makes up

With agriculture bacillus mediated rice callus metaplasia method with pActin::SUN1-mu rice transformation mutant nal1, transformed plant is carried out PCR to be identified, PCR identifies that the primer is above-mentioned P1/P2, the result obtains 23 strain positive plants, be T0 for plant, this positive plant is the plant of transgene SUN1mu, and note is made transfer-gen plant.

Simultaneously in contrast, with in contrast without the rice mutant nal1 of any processing with the plant that changes empty carrier pCAMBIA2300-Actin over to.

4, the phenotype analytical of transfer-gen plant

Observe the phenotype of statistics transfer-gen plant, adjoining tree, wild-type Zhejiang spoke 802 (being called for short zf802).Again the stem height of plant, main spike length degree, main grain number per spike are carried out statistical study.3 repetitions, results averaged ± standard deviation are established in experiment.

The phenotype of transfer-gen plant and adjoining tree is shown in Fig. 2,3,4.Among Fig. 4, every picture group is followed successively by zf802, adjoining tree, transfer-gen plant from left to right.Statistics is as shown in table 1.Show that the transfer-gen plant plant height uprises, blade obviously broadens, color shoals.

The result: plant type changes: ((72.8 ± 5.1cm) obviously uprise the stem height of transfer-gen plant according to plant in 82.5 ± 3.4cm) comparisons; The output phenotype changes: ((20.7 ± 2.3cm) is obviously elongated, and main grain number per spike (208.1 ± 30.1) comparison of transfer-gen plant is according to plant (114.5 ± 30.1) showed increased according to plant in 26.3 ± 1.9cm) comparisons for the main spike length degree of transfer-gen plant.

Adjoining tree refers to rice mutant nal1 without any processing among Fig. 2-4 and in the table 1; Plant that changes empty carrier pCAMBIA2300-Actin over to and coming to the same thing without the rice mutant nal1 of any processing.

Table 1, the statistical study of transfer-gen plant phenotype

	zf802	Adjoining tree	Transfer-gen plant
				Stem height (cm)	81.8±2.4	72.8±5.1	82.5±3.4
Main spike length degree (cm)	24.5±1.6	20.7±1.3	26.3±1.9
				Main grain number per spike	157.4±29.0	114.6±30.1	208.1±30.1

Embodiment 3, gene action mechanism

The blade of the transfer-gen plant among the embodiment 2, adjoining tree and wild-type Zhejiang spoke 802 and the 4th internode that obviously shortens are carried out the paraffin section analysis.

Result such as table 2 and shown in Figure 5.In the rotaring gene plant blade (blade) small bundle (sv) sum be significantly increased in adjoining tree is compared, even surpassed wild-type.Small bundle (sv) number between the two adjacent great vascular bundles (lv) is compared also obviously increase in the rotaring gene plant blade (blade) with adjoining tree simultaneously, thereby causes transfer-gen plant obviously to broaden.Show that the SUN1mu gene may influence the width and the size of blade by the formation of small bundle (sv) in the regulation and control blades (blade), the fascicular generation type of SUN1mu gene pairs also plays important regulation simultaneously.Adjoining tree refers to the rice mutant nal1 without any processing in the table 2; Plant that changes empty carrier pCAMBIA2300-Actin over to and coming to the same thing without the rice mutant nal1 of any processing.

Table 2, transfer-gen plant vascular bundle number

leaf	ZF802	Adjoining tree	Transfer-gen plant
				Leaf wide (cm)	1.7±0.1	1.4±0.1	2.7±0.2
Leaf long (cm)	31.1±6.2	28.3±2.1	47.1±4.8
				The great vascular bundle number	9.6±0.6	9.5±0.8	11.2±0.7
The small bundle number	45.7±3.7	33.3±3.5	66.2±7.3
				Small bundle number between the two adjacent great vascular bundles	4.9±0.4	3.5±0.3	5.9±0.5

Sequence table

＜110〉Inst. of Genetics and Development Biology, CAS

＜120〉a kind of relevant with plant type and/or with output associated protein and encoding gene and application

<160>4

 

<210>1

<211>1375

<212>DNA

＜213〉Oryza paddy rice (Oryza sativa)

<400>1

tgaccccaac?cccaaaccca?ctctactcta?ctgtgcctca?cctcttgcca?ctactatttc 60

tagtagtcgt?gtatcatcat?ttcagatatc?atatcgccac?ctctcgtttt?tttaataata 120

tcagcggcga?gcgagcgaga?tggactcccc?gtcgcctatg?gcggcgcagg?cggccgacct 180

gtcgctgacg?ctggcgccgt?cgggaggggg?tggtggggga?ggaggaggcg?gcggcggtgg 240

tgggtcgtcg?tcggcgtgca?tcgacggcaa?ggacgtgcgg?ctgttcccgt?gcttgttctg 300

caacaagaag?ttcttgaagt?cgcaggcgct?aggcgggcac?cagaacgcgc?acaaagaagg 360

agcggagcat?cgggtggaat?ccctacttct?acatgccgcc?gacgccgcac?cccgccggca 420

atgccgccgc?cgccgccgcg?gcggcgacgc?ccggtgggat?gtcgtccgtc?acgacgccgt 480

ccgggagcta?cggcgtcgtc?ggtggtgccg?ccgccgcggc?ggcggctgtc?gtcggggcta 540

ctgctggcgt?tgggggcgga?ggtggagtgg?gaggggggct?tctcccggcg?cacgcgtacg 600

ccgggcacgg?gtacgccgcg?gtgccgacgt?cgttccccat?cgcgtcgcac?agctcgagcg 660

tggttggctc?cggtgggctg?cagtactacg?ctggtaccga?ctgcggcgcg?gcggcggcgg 720

gtgcggcgaa?gacgacgacg?acgacggcgg?cggcggcggc?gacggccgtg?gcggggagcg 780

agagcggcgt?gcaggtgccc?cggttcgcga?cgcaccagca?ccatctcctg?gcggtggtga 840

gcagcgggcg?cgcgatgctg?gcggcgcccg?accagccggg?cgccgggcgc?gacgacatga 900

tcgacatgct?caactggagg?cgaggctccc?acggccccac?cgcctccgcc?gccgccacca 960

cgccctcccc?ggcaagcacc?accaccacgc?tcaccacctt?cgccagcgcc?gacggcagca 1020

acaacggcga?ggagaacgag?gagctcgacc?tcaacttgag?cctctagctc?ccaccaccac 1080

cacctcctcc?tccgccgccg?ccgccgccgc?cgcgcaatcc?aagaaggcaa?ggtcaatcaa 1140

tcgccatgtt?cttcttctcc?aagctccacc?tactcctctt?ccaattcctc?ctcgtgtgtg 1200

attaatcccc?ctcttcttgc?tgcctgcgta?cgtactcctt?aattaattag?ctcttaggga 1260

cgttaattaa?tctcagttct?tggctctctt?ctcctctcct?ctcctctcct?ctcatctcac 1320

ttgtatgtta?atgttagtac?tccttgtaat?cgatcaatca?gtcctctttt?tttgc 1375

<210>2

<211>353

<212>PRT

＜213〉Oryza paddy rice (Oryza sativa)

<400>2

Met?Asp?Ser?Pro?Ser?Pro?Met?Ala?Ala?Gln?Ala?Ala?Asp?Leu?Ser?Leu

1 5 10 15

Thr?Leu?Ala?Pro?Ser?Gly?Gly?Gly?Gly?Gly?Gly?Gly?Gly?Gly?Gly?Gly

20 25 30

Gly?Gly?Gly?Ser?Ser?Ser?Ala?Cys?Ile?Asp?Gly?Lys?Asp?Val?Arg?Leu

35 40 45

Phe?Pro?Cys?Leu?Phe?Cys?Asn?Lys?Lys?Phe?Leu?Lys?Ser?Gln?Ala?Leu

50 55 60

Gly?Gly?His?Gln?Asn?Ala?His?Lys?Glu?Gly?Ala?Glu?His?Arg?Val?Glu

65 70 75 80

Ser?Leu?Leu?Leu?His?Ala?Ala?Asp?Ala?Ala?Pro?Arg?Arg?Gln?Cys?Arg

85 90 95

Arg?Arg?Arg?Arg?Gly?Gly?Asp?Ala?Arg?Trp?Asp?Val?Val?Arg?His?Asp

100 105 110

Ala?Val?Arg?Glu?Leu?Arg?Arg?Arg?Arg?Trp?Cys?Arg?Arg?Arg?Gly?Gly

115 120 125

Gly?Cys?Arg?Arg?Gly?Tyr?Cys?Trp?Arg?Trp?Gly?Arg?Arg?Trp?Ser?Gly

130 135 140

Arg?Gly?Ala?Ser?Pro?Gly?Ala?Arg?Val?Arg?Arg?Ala?Arg?Val?Arg?Arg

145 150 155 160

Gly?Ala?Asp?Val?Val?Pro?His?Arg?Val?Ala?Gln?Leu?Glu?Arg?Gly?Trp

165 170 175

Leu?Arg?Trp?Ala?Ala?Val?Leu?Arg?Trp?Tyr?Arg?Leu?Arg?Arg?Gly?Gly

180 185 190

Gly?Gly?Cys?Gly?Glu?Asp?Asp?Asp?Asp?Asp?Gly?Gly?Gly?Gly?Gly?Asp

195 200 205

Gly?Arg?Gly?Gly?Glu?Arg?Glu?Arg?Arg?Ala?Gly?Ala?Pro?Val?Arg?Asp

210 215 220

Ala?Pro?Ala?Pro?Ser?Pro?Gly?Gly?Gly?Glu?Gln?Arg?Ala?Arg?Asp?Ala

225 230 235 240

Gly?Gly?Ala?Arg?Pro?Ala?Gly?Arg?Arg?Ala?Arg?Arg?His?Asp?Arg?His

245 250 255

Ala?Gln?Leu?Glu?Ala?Arg?Leu?Pro?Arg?Pro?His?Arg?Leu?Arg?Arg?Arg

260 265 270

His?His?Ala?Leu?Pro?Gly?Lys?His?His?His?His?Ala?His?His?Leu?Arg

275 280 285

Gln?Arg?Arg?Arg?Gln?Gln?Gln?Arg?Arg?Gly?Glu?Arg?Gly?Ala?Arg?Pro

290 295 300

Gln?Leu?Glu?Pro?Leu?Ala?Pro?Thr?Thr?Thr?Thr?Ser?Ser?Ser?Ala?Ala

305 310 315 320

Ala?Ala?Ala?Ala?Ala?Gln?Ser?Lys?Lys?Ala?Arg?Ser?Ile?Asn?Arg?His

325 330 335

Val?Leu?Leu?Leu?Gln?Ala?Pro?Pro?Thr?Pro?Leu?Pro?Ile?Pro?Pro?Arg

340 345 350

Val

 

<210>3

<211>20

<212>DNA

＜213〉artificial sequence

<220>

<223>

<400>3

atggactccc?cgtcgcctat 20

<210>4

<211>22

<212>DNA

＜213〉artificial sequence

<220>

<223>

<400>4

tcacacacga?ggaggaattg?ga 22

Claims (10)

1. protein, be following a) or b) protein:

A) protein of forming by the aminoacid sequence shown in the SEQ ID NO:2;

B) with the aminoacid sequence shown in the SEQ ID NO:2 through the replacement of one or several amino-acid residue and/or disappearance and/or interpolation and relevant with the plant plant type and/or be correlated with output by a) deutero-protein.

2. the described proteinic encoding gene of claim 1.

3. encoding gene according to claim 2 is characterized in that: described proteic encoding gene is following 1), 2), 3) or 4) gene:

1) its nucleotide sequence is from dna molecular shown in the Nucleotide of 5 ' terminal 140-1201 position among the SEQ ID NO:1;

2) its nucleotide sequence is a dna molecular shown in the SEQ ID NO:1;

3) under stringent condition with 1) or 2) the dna sequence dna hybridization that limits and the dna molecular of encoding said proteins;

4) with 1) or 2) dna sequence dna that limits has the homology more than 90% and the dna molecular of encoding said proteins.

4. amplification claim 2 or 3 described full length genes or its any segmental primer are right; Or a primer sequence of described primer centering is shown in SEQ ID NO:3, and another primer sequence of described primer centering is shown in SEQ ID NO:4.

5. the recombinant vectors, reorganization bacterium, transgenic cell line or the expression cassette that contain claim 2 or 3 described encoding genes;

Or described recombinant vectors is that the multiple clone site that claim 2 or 3 described encoding genes insert carrier pCAMBIA2300-Actin is obtained.

6. claim 2 or the 3 described encoding genes application in improveing plant plant type and/or improvement output phenotype and/or raising output, or the application of the described protein of claim 1 in improvement plant plant type and/or improvement output phenotype and/or raising output.

7. a method that improves plant plant type and/or improvement output phenotype and/or improve output is following 1) or 2) or 3) described:

1) in the plant that sets out, imports claim 2 or 3 described encoding genes, obtain comparing the purpose plant of plant type improvement with the plant that sets out;

2) in the plant that sets out, import claim 2 or 3 described encoding genes, obtain comparing the purpose plant of output phenotype improvement with the plant that sets out;

3) in the plant that sets out, import claim 2 or 3 described encoding genes, obtain comparing the purpose plant of output raising with the plant that sets out.

8. method according to claim 7 is characterized in that: claim 2 or 3 described encoding genes import by recombinant vectors described in the claim 5.

9. application according to claim 6 or claim 7 or 8 described methods, it is characterized in that: described plant type improvement is wider than the plant that sets out for the tall and big plant leaf in the plant that sets out, purpose plant of plant stem of purpose plant, and/or the plant leaf of purpose plant is grown up in the plant that sets out;

The improvement of described output phenotype for the plant master spike length of purpose plant greater than the plant master grain number per spike of set out plant and/or purpose plant more than the plant that sets out.

10. arbitrary described method among application according to claim 6 or the claim 7-9 is characterized in that: the described plant that sets out is a monocotyledons; Described monocotyledons is a paddy rice; Described paddy rice is rice mutant nall.

Images